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Uptake of 2-NBDG as a method to monitor therapy response in breast cancer cell lines

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Abstract

This study quantifies uptake of a fluorescent glucose analog, (2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose) (2-NBDG), in a large panel of breast cancer cells and demonstrates potential to monitor changes in glycolysis caused by anticancer and endocrine therapies. Expressions of glucose transporter (GLUT 1) and hexokinase (HK I), which phosphorylates 2-NBDG, were measured via western blot in two normal mammary epithelial and eight breast cancer cell lines of varying biological subtype. Fluorescence intensity of each cell line labeled with 100 μM 2-NBDG for 20 min or unlabeled control was quantified. A subset of cancer cells was treated with anticancer and endocrine therapies, and 2-NBDG fluorescence changes were measured. Expression of GLUT 1 was necessary for uptake of 2-NBDG, as demonstrated by lack of 2-NBDG uptake in normal human mammary epithelial cells (HMECs). GLUT 1 expression and 2-NBDG uptake was ubiquitous among all breast cancer lines. Reduction and stimulation of 2-NBDG uptake was demonstrated by perturbation with anticancer agents, lonidamine (LND), and α-cyano-hydroxycinnamate (α-Cinn), respectively. LND directly inhibits HK and significantly reduced 2-NBDG fluorescence in a subset of two breast cancer cell lines. Conversely, when cells were treated with α-Cinn, a drug used to increase glycolysis, 2-NBDG uptake was increased. Furthermore, tamoxifen (tam), a common endocrine therapy, was administered to estrogen receptor positive and negative (ER+/−) breast cells and demonstrated a decreased 2-NBDG uptake in ER+ cells, reflecting a decrease in glycolysis. Results indicate that 2-NBDG uptake can be used to measure changes in glycolysis and has potential for use in early drug development.

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Acknowledgments

The authors would like to acknowledge the support of the NIH Imaging Training Grant and DOD grant W81XWH-05-1-0363. Also, the Seewaldt lab has been especially helpful and is greatly appreciated for training SM in the use of molecular assays and for providing access to appropriate laboratory equipment.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Duke University, 136 Hudson Hall, Campus, Box 90281, Durham, NC, 27708-0281, USA

    Stacy R. Millon, J. Quincy Brown, Anita Raheja & Nirmala Ramanujam

  2. Department of Medicine, Duke University, Durham, NC, USA

    Julie H. Ostrander & Victoria L. Seewaldt

Authors
  1. Stacy R. Millon
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  2. Julie H. Ostrander
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  3. J. Quincy Brown
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  4. Anita Raheja
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  5. Victoria L. Seewaldt
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  6. Nirmala Ramanujam
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Correspondence to Stacy R. Millon.

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Millon, S.R., Ostrander, J.H., Brown, J.Q. et al. Uptake of 2-NBDG as a method to monitor therapy response in breast cancer cell lines. Breast Cancer Res Treat 126, 55–62 (2011). https://doi.org/10.1007/s10549-010-0884-1

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  • DOI: https://doi.org/10.1007/s10549-010-0884-1

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